Page 54 - 《中国药房》2020年22期

尼可地尔对肺动脉平滑肌细胞增殖、迁移能力和Hippo/YAP信号

通路的影响 Δ

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陈枫楠 ，郎廷元 ，吴常裕 ，俞晓军 ，石新木 ，沈 凯 ，杨海燕 （1.舟山市妇女儿童医院内二科，浙江 舟山
316004；2.重庆大学附属肿瘤医院妇科肿瘤中心，重庆 400030；3.浙江省舟山医院心内科，浙江 舟山
316021）
中图分类号 R361 .3；R965 文献标志码 A 文章编号 1001-0408（2020）22-2736-05
+
DOI 10.6039/j.issn.1001-0408.2020.22.09
摘 要 目的：体外评价尼可地尔对肺动脉平滑肌细胞（PASMCs）增殖、迁移能力和Hippo/YAP信号通路的影响。方法：将人原代
PASMCs分为正常对照组、模型组和尼可地尔低、中、高浓度组（50、100、200 μmol/L），每组设3个复孔。除正常对照组外，其余各
组细胞均接种在凝胶包被的培养板上，模拟肺动脉高压环境，以建立肺动脉硬化（AS）细胞模型；随后，各药物组加入相应药物，正
常对照组和模型组加入同等体积生理盐水，培养48 h。采用CCK-8法检测各组细胞的增殖能力（以光密度计），Transwell方法检测
细胞迁移能力，实时定量聚合酶链式反应技术检测细胞中 YAP 靶基因子[结缔组织生长因子（CTGF）、双向调节蛋白（AREG）]
mRNA的表达水平，Western blotting法检测细胞中CTGF、AREG蛋白的表达水平。结果：与正常对照组比较，模型组细胞的光密
度显著升高，每视野迁移细胞数显著增加，细胞中CTGF、AREG mRNA及蛋白的表达水平均显著增强（P＜0.01）。与模型组比较，
尼可地尔低、中、高浓度组细胞的光密度、每视野迁移细胞数和细胞中CTGF、AREG mRNA及蛋白的表达水平均显著降低，且成
浓度依赖性（P＜0.05或P＜0.01）。结论：尼可地尔可抑制AS模型PASMCs的增殖和迁移，其作用机制可能与抑制Hippo/YAP信
号通路有关。
关键词 尼可地尔；肺动脉高压；肺动脉硬化；肺动脉平滑肌细胞；Hippo/YAP信号通路

Effects of Nicorandil on the Proliferation， Migration Ability and Hippo/YAP Signaling Pathway of
Pulmonary Artery Smooth Muscle Cells
CHEN Fengnan ，LANG Tingyuan ，WU Changyu ，YU Xiaojun ，SHI Xinmu ，SHEN Kai ，YANG Haiyan 1
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（1.Dept. Two of Internal Medicine，Zhoushan Women and Children Hospital，Zhejiang Zhoushan 316004，
China；2. Gynecological Oncology Center，Chongqing University Cancer Hospital，Chongqing 400030，China；
3. Dept. of Cardiology，Zhoushan Hospital of Zhejiang Province，Zhejiang Zhoushan 316021，China）
ABSTRACT OBJECTIVE：To evaluate the effects of nicorandil on the proliferation，migration ability and Hippo/YAP signaling
pathway of pulmonary artery smooth muscle cells（PASMCs）. METHODS：Human primary PASMCs were divided into normal
control group，model group，nicorandil low，medium and high concentration groups（50，100，200 μmol/L），with 3 holes in each
group. In addition to the normal control group，the rest of the cells were inoculated on the gel coated medium to simulate the
pulmonary hypertension environment，so as to establish AS cell model. Then，each drug group was added with corresponding
drugs，and the normal control group and model group were added with the same volume of normal saline，and cultured for 48 h.
CCK-8 assay and Transwell assay were used for the examination of cell proliferation （by light density） and migration ability，
respectively. mRNA expression of YAP target factors（CTGF and AREG）were examined by qRT-PCR. Western blotting assay was
used to detect the protein expression of CTGF and AREG. RESULTS：Compared with normal control group，light density of cells
was increased significantly in model group；the number of migration cells per field of view increased significantly；mRNA and
protein expression of CTGF and AREG were significantly increased（P＜0.01）. Compared with model group，light density，the
number of migration cells per field of view，mRNA and protein expression of CTGF and AREG in nicorandil low，medium and
high concentration groups were decreased significantly， in concentration-dependent manner （P＜0.05 or P＜0.01）.
CONCLUSIONS：Nicorandil can inhibit the proliferation and
Δ 基金项目：重庆市科技局技术创新与应用发展面上项目（No.
migration of PASMCs in AS model，the mechanism of which
cstc2019jscx-msxmX0174）；浙 江 省 医 药 卫 生 科 技 计 划 面 上 项 目
may be associated with the Hippo/YAP signaling pathway.
（No.2018241273）
KEYWORDS Nicorandil；Pulmonary arterial hypertension；
＊住院医师。研究方向：心血管。电话：0580-2065131。E-mail：
343070302@qq.com AS； Pulmonary arterial smooth muscle cells； Hippo/YAP
# 通信作者：主任医师。研究方向：肺动脉高压。电话：0580- signaling pathway
2065131。E-mail：yanghaiyan1975@163.com

·2736 · China Pharmacy 2020 Vol. 31 No. 22 中国药房 2020年第31卷第22期